Overproduction of medicinal ergot alkaloids based on a fungal platform

• Published in: Metabolic engineering 2022-01, Vol.69, p.198-208
• Main Authors: Yao, Yongpeng, Wang, Wei, Shi, Wenyu, Yan, Rui, Zhang, Jun, Wei, Guangzheng, Liu, Ling, Che, Yongsheng, An, Chunyan, Gao, Shu-Shan
• Format: Article
• Language: English
• Subjects: Aspergillus nidulans; Biosynthetic Pathways - genetics; Claviceps - genetics; Claviceps - metabolism; Cytochrome P-450 Enzyme System - genetics; Cytochrome P-450 Enzyme System - metabolism; Ergot alkaloids; Ergot Alkaloids - genetics; Ergot Alkaloids - metabolism; Heterologous production; P450 electron-transfer engineering; Pathway refactoring; Saccharomyces cerevisiae - metabolism
• ISSN: 1096-7176; 1096-7184
• DOI: 10.1016/j.ymben.2021.12.002

Privileged ergot alkaloids (EAs) produced by the fungal genus Claviceps are used to treat a wide range of diseases. However, their use and research have been hampered by the challenging genetic engineering of Claviceps. Here we systematically refactored and rationally engineered the EA biosynthetic pathway in heterologous host Aspergillus nidulans by using a Fungal-Yeast-Shuttle-Vector protocol. The obtained strains allowed the production of diverse EAs and related intermediates, including prechanoclavine (PCC, 333.8 mg/L), chanoclavine (CC, 241.0 mg/L), agroclavine (AC, 78.7 mg/L), and festuclavine (FC, 99.2 mg/L), etc. This fungal platform also enabled the access to the methyl-oxidized EAs (MOEAs), including elymoclavine (EC), lysergic acid (LA), dihydroelysergol (DHLG), and dihydrolysergic acid (DHLA), by overexpressing a P450 enzyme CloA. Furthermore, by optimizing the P450 electron transfer (ET) pathway and using multi-copy of cloA, the titers of EC and DHLG have been improved by 17.3- and 9.4-fold, respectively. Beyond our demonstration of A. nidulans as a robust platform for EA overproduction, our study offers a proof of concept for engineering the eukaryotic P450s-contained biosynthetic pathways in a filamentous fungal host. [Display omitted] •The heterologous refactoring of EA BGCs enabled A. nidulans to produce diverse EAs.•This platform produced 78.7 mg/L of agroclavine and 99.2 mg/L of festuclavine.•This platform allowed to produce lysergic acid and dihydrolysergic acid.•Optimization of ET system of CloA improved the production of elymoclavine and dihydroelysergol.